Molecular Analysis of Single-Nucleotide Polymorphisms of TRAF5 Gene in Patients with Behçet's Disease from the Azeri Population of Northwest Iran

Research Article | DOI: https://doi.org/DOI: 10.31579/2690-4861/148

Molecular Analysis of Single-Nucleotide Polymorphisms of TRAF5 Gene in Patients with Behçet's Disease from the Azeri Population of Northwest Iran

  • Reza Hosseinzadeh 1
  • Leila Saremi 2
  • Mohammad Ebrahim Ghaffari 3
  • Sepideh Babaniamansour 4
  • Mohammad Khalaj-Kondori * ID 5

1MS, Department of Genetic, East Azarbayjan Sciences and Research Branch, Islamic Azad University, Tabriz, Iran.

2PhD, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

3PhD in Biostatistics, Dental Sciences, Research Center, Faculty of Dentistry, Guilan University of MedicalSciences, Rasht, IRAN.

4MD, Department of Pathology, Rogel Cancer Center and Center for RNA Biomedicine, University of Michigan, Ann Arbor, MI 48109, USA.

5PhD, Department of Animal Biology Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.

*Corresponding Author: Mohammad Khalaj-Kondori, Associate Professor of Molecular Genetics, Department of Animal Biology Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.

Citation: R Hosseinzadeh, L Saremi, Mohammad E Ghaffari, S Babaniamansour, Mohammad K Kondori. (2021) Molecular Analysis of Single-Nucleotide Polymorphisms of TRAF5 Gene in Patients with Behçet's Disease from the Azeri Population of Northwest Iran. International Journal of Clinical Case Reports and Reviews. 8(2); DOI:10.31579/2690-4861/148

Copyright: © 2021 Mohammad Khalaj-Kondori, This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Received: 17 June 2021 | Accepted: 11 August 2021 | Published: 20 August 2021

Keywords: behçet syndrome; genetic polymorphism; human traf5 protein; polymerase chain reaction

Abstract

Background: Behçet's disease (BD) is a chronic inflammatory disease with unknown causes. The geographical distribution is mostly consistent with the historic Silk Road. The role of tumor necrosis factor receptor-associated factor 5 (TRAF5) gene in the inflammation signaling pathway leads more attention to the potency of different polymorphisms of the TRAF5 gene, in the development of BD.

Methods: This was a case-control study conducted among the Azeri Turk ethnic group (50 BD patients and 50 persons with no history of autoimmune disease), Tabriz, Iran. Four different TRAF5 gene polymorphisms were assessed via the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism technique. Data were analyzed by SPSS Version 20.

Results: Genotypic distributions of four studied Single nucleotide polymorphisms (SNPs) had no significant difference between case and control groups (rs6540679 (p=0.50), rs12569232 (p=0.86), rs10863888 (p=0.14), and rs7514863 (p=0.24).  There were also found no significant differences in SNP allelic frequencies between cases and controls (all 

Background

Behçet's disease (BD) is a multifactorial inflammatory and systemic uveitis autoimmune disease, which is associated with recurrent skin lesions, oral aphthae, and genital ulcers with unknown causes [1-6]. The highest incidence of BD belongs to Turkey (80-370 patients in 100,000 persons), whereas it has a low rate in North America (0.2 patients in 100,000 people) [7]. The onset age of the disease is usually between 25 and 30 years old [8]. The disease is equally distributed between genders, but the previous studies reported higher severity in males than females for unknown reasons.  Although numerous studies have been done, the main causes of the BD are still unclear [9].  

Recently, the role of various active oxygen derivatives (free radicals) including superoxide, hydroxyl peroxide, hydroxyl radicals, and nitric oxide as well as the activity of antioxidant enzymes (dismutase oxide and glutathione peroxidase) in the pathogenesis of BD were examined. These elements have been considered as transcription activators of pro-inflammatory cytokines [10, 11]. Additionally, a wide range of mutations and polymorphisms in association with BD patients have been reported, such as human leukocyte antigen (HLA-B) serotype 5, chemokine receptor 5, heat shock protein 60, anti-tumor necrosis factor (TNF) α, Interleukin-6 (IL-6), and Interferon-gamma (IFN-γ), Cluster of differentiation 40 (CD40), signal transducer and activator of transcription 3 (STAT3), STAT4, and Janus kinase 2 (JAK2) [9, 12], Intercellular adhesion molecule-1, small ubiquitin-like modifier 4, IL-23R-IL12RB2, IL10, Monocyte chemoattractant protein-1, and Fc receptor-like 3 [13].

Moreover, dominant immune responses in the patients are considered important factors in its pathogenesis. The physicians choose Corticosteroids, Colchicine, Azathioprine, Cyclosporine, Thalidomide, Cyclophosphamide, Interferon-α, or TNFα inhibitors as the BD treatment based on the severity of the organ involvement and clinical symptoms [14].

Tumor necrosis factor receptor-associated factors (TRAFs) are an intra-cellular family of adaptor proteins that share a common structural domain at the C-terminus and directly attach to TNF receptors (TNF-R) [15, 16]. There are seven known types of TRAF family (TRAF1 to 7) in mammals [17]. All TRAFs, except TRAF1, consist of an N-terminal ring finger domain followed by the various number of zinc fingers [16, 18]. The ring finger consists of one histidine residue and seven cysteine residues forming a single folded domain binding two zinc ions. In ubiquitin ligases, ring finger motifs are vital for ubiquitin molecule transfer to substrates or ring finger motifs themselves. TRAFs are related to various diseases such as immunodeficiency, cancer pathogenesis, autoimmune, and neurodegenerative [19].

The TRAF5 identified as a CD40 binding protein and a lymphotoxin-b receptor-interacting protein (LTBR), locates on the humans’ chromosome 1q32.2. (Figure 1)  It significantly expresses in the thymus gland, lungs, spleen, kidneys as well as peripheral blood cells and plays an important role in cell signaling [20]. TRAF5 using the yeast two-hybrid system, and LBR, by PCR using degenerate primers corresponding to conserved amino acids in the TRAF domain of TRAF1-3 [21].

Figure 1: Chromosomal location of TRAF5.

Single nucleotide polymorphism (SNP) in TARF is related to autoimmune diseases such as lupus erythematosus, rheumatoid arthritis (RA) [22-25]. Xiang et al. demonstrated that TRAF5 polymorphisms are related to susceptibility to autoimmune diseases such as RA. An SNP (rs7514863) upstream of TRAF5 that affects a transcriptional factor binding site is associated with RA [26]. A major dominant mutation in TRAF3 was reported in a young child with a herpes simplex virus (HSV1) encephalitis history [27]. A TRAF6 heterozygous mutation has been recently reported in a patient with Hypohidrotic Ectodermal Dysplasia [28]. In 2014, the association between rs10863888 and BD was observed in a Chinese population [20]. 

We selected four SNPs (rs1256932, rs6540679, rs10863888, and rs7514863) for the investigation of TRAF5 polymorphisms based on reports in psoriasis and RA [29], linkage disequilibrium patterns in Han Chinese, and Japanese data in the Hap Map database.

Methods

Study design

This was a case-control study conducted at the Molecular-Medical Genetic department of Islamic Azad University East Azarbaijan Science and Research Branch, Tabriz, Iran. The executive protocol of the study was conducted under the Declaration of Helsinki (seventh revision 2013). All participants had written consent before the study.

Participants

In this study, the case group included 50 BD patients from the Azeri Turk ethnic group, diagnosed by a rheumatologist (26 males and 24 females in each group). The control group included 50 persons with neither history of autoimmune disease nor a family history of autoimmune disease.

Genomic deoxyribonucleic acid (DNA) extraction using the Salting out method

A venous blood sample (5 cc) was obtained from all participants and collected in Ethylenediaminetetraacetic acid (EDTA, Merck Company, Germany)-containing falcon (SARSTEDT Company, Germany). The samples were shaken slowly to mix and prevent the formation of blood clots. DNA was extracted from blood samples based on the Nasiri et. al, methods [30]. The quantity and quality of extracted DNA were determined using spectrophotometry and agarose gel (Kosar Company, Iran) electrophoresis, respectively. 

Polymerase Chain Reaction (PCR)

PCR (ABI Company, USA) was performed to amplify specific DNA sequences using specific primers (manufactured by Biorad Company, Germany). Amplification conditions for PCR reactions were as follows: one cycle at 95 °C for 5 min, 30 cycles at 94 °C for 25 s, 63 °C for 20 s and 72 °C for 20 s, followed by a final extension at 72 °C for 5 min. PCR products were electrophoresed on agarose gels (2%), stained with the ethidium bromide (manufactured by CinnaGen, Iran), and documented.

Digestion of PCR-products with restriction enzymes (PCR -RFLP)

PCR-Restriction Fragment Length Polymorphism (RFLP) is commonly used to examine the various alleles of a gene in a population. The sequence diversity of the gene corresponding alleles creates different digestion locations for the restriction enzyme, and segments with different lengths are obtained. Induced mutations in the detection site of restriction enzymes (TRUII-SacI-TSP509I-APOI-hinfI, manufactured by Fermentas company, Germany) and resulted polymorphism can be detected by its RFLP pattern. After amplification of the mentioned DNA segments, PCR products were digested using restriction enzymes. Each reaction tubes contain 9 µl ddH2O, 1 µl buffer 10 x, 0.5 µl restriction enzyme, and 5 µl PCR product, then incubated overnight at 37 °C. The position information of each of the polymorphisms, restriction enzymes, and DNA fragment size is listed in table 1. Finally, the incubated reactions were electrophoresed on agarose 3% and documented. 

Table 1: Primers, enzymes, and the size of the digested fragments using digestion enzymes

Statistical analysis

A Chi-square test with continuity correction was used to examine the Hardy-Weinberg equilibrium. After it was found that Hardy-Weinberg equilibrium is established in all polymorphisms, Chi-square and Fisher's exact tests were used to compare the frequency of alleles and genotypes between case and control groups. To perform calculations, R version 3.4.3 and SPSS version 20 (SPSS Inc., Chicago, IL, USA) were used to analyze the data. P<0>

Results

The present study was conducted among 100 participants. The mean age of participants in the case and control groups were 34.02±7.39 (range 11 to 65) and 34.42±8.27 (range 21 to 76) years, respectively. Patients in the case group had no family history of BD. The frequency of the positive Pathergy test in the case and control groups were 14(28%) and zero, respectively. The clinical characteristic of the case group is shown in table 2.

Table 2: Clinical characteristics of case group.

In this study, four polymorphisms (rs12569232, rs6540679, rs7514863, and rs10863888) of the TRAF5 gene were evaluated. To investigate the polymorphism of rs6540679 (SNP: rs6540679 Chr.1: 211491152) a segment about 233 bp was amplified using PCR followed by digestion using the Xmil (ACC I) enzyme, which PCR products that contain allele (G) was not digested but a product with allele (A) digested and fragments with 100 and 133 bp were produced. The people who were heterozygous for the given site showed three bands with lengths of 133, 100, and 233 bp. (Figure 2).

Figure 2. Polymorphism of rs6540679.Lane C: Ladder 100 bp, Lane A, E, F and G: GG genotype (rs6540679) (233 bp), lane D: AA genotype (rs6540679) (100, 133 bp), Lane B: AG genotype (rs6540679) (100, 133, 233 bp).

To assess the polymorphism of rs12569232 (SNP: rs12569232 Chr.1: 211553064) a segment, about 305 bp was amplified using PCR followed by digestion using the Mva I (BstNI) enzyme, which PCR products contain allele (C) was not digested but a product with allele (G) digested and fragments 220 and 85 bp were produced. The people who were heterozygous for the given site showed three segments with lengths of 220, 85, and 305 bp. (Figure 3).

Figure 3. Polymorphism of rs12569232.Lane A: Ladder 100 bp, Lane B and E: CC genotype (rs12569232) (305 bp), Lane D: GG genotype (rs12569232) (85, 220 bp), Lane C: CG genotype (rs12569232) (85, 220, 305 bp).

To study the polymorphism of rs10863888 (SNP: rs10863888 Chr.1: 211502769) a fragment about 311 bp was amplified using PCR followed by digestion using the Mae III enzyme, which PCR products that contain allele (A) digested, but allele (G) was not digested and fragments with 288 and 24 bp produced. The heterozygous people for the given site showed three bands with lengths of 288, 24 and, 311 bp. (Figure 4).

Figure 4. Polymorphism of rs10863888. Lane E: Ladder 100 bp.

To assess the polymorphism of rs7514863 a fragment of about 254 bp was amplified and followed by digestion using the Tsp 451 enzyme, which PCR products contain allele (A) digested but a product with allele (G) was not digested and fragments 231 and 23 bp were produced. The heterozygous individuals for the given site showed three segments with lengths of 254, 231, and 23 bp. (Figure 5).

Figure 5. Polymorphism of rs7514863. Lane D: Ladder 100 bp.

Tables 3 and 4 show the frequency of allele and genotype in four different polymorphisms of the TRAF5 gene between case and control groups. The results showed no significant difference for rs6540679 polymorphism (P=0.92) (OR=0.895, 0.41-1.94), rs12569232 (P=0.75, OR=1.17, 0.61-2.24), rs10863888 (P=0.088; OR= 1.76, 0.97-3.21) and rs7514863 (P=0.65, OR= 0.674, 0.23-1.967) between case and control groups. (Table 5).

Table 3: The frequency of each allele between case and control groups
Table 4: The frequency of each genotype between case and control groups.
Table 5: The frequency of gene polymorphisms (allele and genotype)

Discussion

In this study, the relationship between common polymorphisms of the TRAF5 gene and BD in the Azari-Iranian population was studied. Although the cause of BD has not been fully characterized, however, it seems that genetic and environmental factors contribute to the disease. For example, HLA-B51 is commonly found in BD patients around the Silk Road. 

The results of this study showed that there was no relationship between polymorphisms of rs6540679, rs12569232, rs10863888, and rs7514863 with the susceptibility to the BD.

The relationship between autoimmune diseases like BD and the TRAF5 gene polymorphisms was assessed in the previous studies but they showed contradictory results. Potter et al. conducted a study among 351 BD patients and 368 persons (as a control group) in England. They examined 44 markers of the TRAF gene such as rs7514863 and showed a strong relationship between the TRAF5 gene polymorphism (rs7514863) and RA. Potter suggested that this marker can be a candidate polymorphism for RA [26].

Xiang et al. studied markers of rs12569232, rs6540679,and rs10863888 in 450 acute anterior uveitis autoimmune patients, 458 pediatric uveitis patients, and 1601 persons as a control group in China. There was a significant relationship between polymorphism of rs12569232 and disease, but no significant relationship was observed for rs6540679 and rs10863888[31].

In the study on 789 BD and 940 patients suffering Vogt-Koyanagi-Harada Syndrome in China, rs10563888 marker was significantly associated with BD, while there was no significant association for rs6540679 and rs12569232 [20]. Studies stated that TNF-α and MEFV polymorphisms had a significant association with BD in Iranian Azeri Turkish [3233]. High expression of JAK1 and TLR2 in patients with BD confirmed the important role of this gene in its development [34].

A study among BD patients in China showed that AG and GG alleles of rs10863888 polymorphism had a significant difference. We thought that if the number of patients and the control group considered on a larger scale the significance of this polymorphism would be very high. Although the P-value obtained for rs10863888 polymorphism in this study is 0.088, confirms our hypothesis that this polymorphism may be meaningful in the case of a larger patient and control community. 

Differences in the results of various studies on the association between the TRAF5 gene polymorphism and BD can be justified for several reasons. First, this difference can be due to the genetic variation of studied populations in various studies, which indicates the presence of different polymorphism levels in different populations. On the other hand, this inconsistency could be due to the low number of surveyed people and used techniques. Thus, it is recommended to carry out more extensive studies with more samples and more precise techniques, including sequencing of the fragments using DNA sequencing techniques.

Conclusion

Although the role of the TRAF5 gene polymorphisms in inflammatory diseases has been proved in many investigations, Our study did not show a significant relationship between the TRAF5 gene polymorphisms and BD.

List of abbreviations

BD: Behçet's Disease; TRAF: Tumor Necrosis Factor Receptor-Associated Factor; SPSS: Statistical Package for the Social Sciences; SNP: Single Nucleotide Polymorphism; HLA: Human Leukocyte Antigen; TNF: Tumor Necrosis Factor; IL: Interleukin; IFN: Interferon; CD: Cluster of Differentiation; STAT: Signal Transducer and Activator of Transcription; JAK: Janus Kinase; LTBR: Lymphotoxin-b Receptor-Interacting Protein; RA: Rheumatoid Arthritis; HSV: Herpes Simplex Virus; DNA: Deoxyribonucleic Acid; EDTA: Ethylenediaminetetraacetic Acid; PCR: Polymerase Chain Reaction; RFLP: Restriction Fragment Length Polymorphism.

Declaration

Ethic approval: The implementation of the project was approved by the ethics committee of Islamic Azad University East Azarbaijan Science.

Consent to participate: Informed consent was obtained from all the participants prior to the study.

Consent for publication: Not applicable.

Availability of data and material: Data sharing is available upon the author's request.

Funding: This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sector.

Competing Interests: On behalf of all authors, the corresponding author states that there is no conflict of interest.

Authors’ contributions

Leila Saremi, Reza Hosseinzadeh and Mohammad Khalaj-Kondori were in charge of samples collection, experimental design and PCR. Leila Saremi, Mohammad Khalaj-Kondori, and Mohammad Ebrahim Ghaffari contributed equally performed and designed experiments, analyzed data, and generated figures. Reza Hosseinzadeh, and Sepideh Babaniamansour were in charge of idea and concept of the paper. Mohammad Ebrahim Ghaffari, Sepideh Babaniamansour and Mohammad Khalaj-Kondori assisted in supervision and generated figures and edited the manuscript. All authors read and approved the final manuscript.

Acknowledgements

We thank Dr. Aziz Khorrami for his excellent technical assistance and contributions in this study.

References

Clearly Auctoresonline and particularly Psychology and Mental Health Care Journal is dedicated to improving health care services for individuals and populations. The editorial boards' ability to efficiently recognize and share the global importance of health literacy with a variety of stakeholders. Auctoresonline publishing platform can be used to facilitate of optimal client-based services and should be added to health care professionals' repertoire of evidence-based health care resources.

img

Virginia E. Koenig

Journal of Clinical Cardiology and Cardiovascular Intervention The submission and review process was adequate. However I think that the publication total value should have been enlightened in early fases. Thank you for all.

img

Delcio G Silva Junior

Journal of Women Health Care and Issues By the present mail, I want to say thank to you and tour colleagues for facilitating my published article. Specially thank you for the peer review process, support from the editorial office. I appreciate positively the quality of your journal.

img

Ziemlé Clément Méda

Journal of Clinical Research and Reports I would be very delighted to submit my testimonial regarding the reviewer board and the editorial office. The reviewer board were accurate and helpful regarding any modifications for my manuscript. And the editorial office were very helpful and supportive in contacting and monitoring with any update and offering help. It was my pleasure to contribute with your promising Journal and I am looking forward for more collaboration.

img

Mina Sherif Soliman Georgy

We would like to thank the Journal of Thoracic Disease and Cardiothoracic Surgery because of the services they provided us for our articles. The peer-review process was done in a very excellent time manner, and the opinions of the reviewers helped us to improve our manuscript further. The editorial office had an outstanding correspondence with us and guided us in many ways. During a hard time of the pandemic that is affecting every one of us tremendously, the editorial office helped us make everything easier for publishing scientific work. Hope for a more scientific relationship with your Journal.

img

Layla Shojaie

The peer-review process which consisted high quality queries on the paper. I did answer six reviewers’ questions and comments before the paper was accepted. The support from the editorial office is excellent.

img

Sing-yung Wu

Journal of Neuroscience and Neurological Surgery. I had the experience of publishing a research article recently. The whole process was simple from submission to publication. The reviewers made specific and valuable recommendations and corrections that improved the quality of my publication. I strongly recommend this Journal.

img

Orlando Villarreal

Dr. Katarzyna Byczkowska My testimonial covering: "The peer review process is quick and effective. The support from the editorial office is very professional and friendly. Quality of the Clinical Cardiology and Cardiovascular Interventions is scientific and publishes ground-breaking research on cardiology that is useful for other professionals in the field.

img

Katarzyna Byczkowska

Thank you most sincerely, with regard to the support you have given in relation to the reviewing process and the processing of my article entitled "Large Cell Neuroendocrine Carcinoma of The Prostate Gland: A Review and Update" for publication in your esteemed Journal, Journal of Cancer Research and Cellular Therapeutics". The editorial team has been very supportive.

img

Anthony Kodzo-Grey Venyo

Testimony of Journal of Clinical Otorhinolaryngology: work with your Reviews has been a educational and constructive experience. The editorial office were very helpful and supportive. It was a pleasure to contribute to your Journal.

img

Pedro Marques Gomes

Dr. Bernard Terkimbi Utoo, I am happy to publish my scientific work in Journal of Women Health Care and Issues (JWHCI). The manuscript submission was seamless and peer review process was top notch. I was amazed that 4 reviewers worked on the manuscript which made it a highly technical, standard and excellent quality paper. I appreciate the format and consideration for the APC as well as the speed of publication. It is my pleasure to continue with this scientific relationship with the esteem JWHCI.

img

Bernard Terkimbi Utoo

This is an acknowledgment for peer reviewers, editorial board of Journal of Clinical Research and Reports. They show a lot of consideration for us as publishers for our research article “Evaluation of the different factors associated with side effects of COVID-19 vaccination on medical students, Mutah university, Al-Karak, Jordan”, in a very professional and easy way. This journal is one of outstanding medical journal.

img

Prof Sherif W Mansour

Dear Hao Jiang, to Journal of Nutrition and Food Processing We greatly appreciate the efficient, professional and rapid processing of our paper by your team. If there is anything else we should do, please do not hesitate to let us know. On behalf of my co-authors, we would like to express our great appreciation to editor and reviewers.

img

Hao Jiang

As an author who has recently published in the journal "Brain and Neurological Disorders". I am delighted to provide a testimonial on the peer review process, editorial office support, and the overall quality of the journal. The peer review process at Brain and Neurological Disorders is rigorous and meticulous, ensuring that only high-quality, evidence-based research is published. The reviewers are experts in their fields, and their comments and suggestions were constructive and helped improve the quality of my manuscript. The review process was timely and efficient, with clear communication from the editorial office at each stage. The support from the editorial office was exceptional throughout the entire process. The editorial staff was responsive, professional, and always willing to help. They provided valuable guidance on formatting, structure, and ethical considerations, making the submission process seamless. Moreover, they kept me informed about the status of my manuscript and provided timely updates, which made the process less stressful. The journal Brain and Neurological Disorders is of the highest quality, with a strong focus on publishing cutting-edge research in the field of neurology. The articles published in this journal are well-researched, rigorously peer-reviewed, and written by experts in the field. The journal maintains high standards, ensuring that readers are provided with the most up-to-date and reliable information on brain and neurological disorders. In conclusion, I had a wonderful experience publishing in Brain and Neurological Disorders. The peer review process was thorough, the editorial office provided exceptional support, and the journal's quality is second to none. I would highly recommend this journal to any researcher working in the field of neurology and brain disorders.

img

Dr Shiming Tang

Dear Agrippa Hilda, Journal of Neuroscience and Neurological Surgery, Editorial Coordinator, I trust this message finds you well. I want to extend my appreciation for considering my article for publication in your esteemed journal. I am pleased to provide a testimonial regarding the peer review process and the support received from your editorial office. The peer review process for my paper was carried out in a highly professional and thorough manner. The feedback and comments provided by the authors were constructive and very useful in improving the quality of the manuscript. This rigorous assessment process undoubtedly contributes to the high standards maintained by your journal.

img

Raed Mualem

International Journal of Clinical Case Reports and Reviews. I strongly recommend to consider submitting your work to this high-quality journal. The support and availability of the Editorial staff is outstanding and the review process was both efficient and rigorous.

img

Andreas Filippaios

Thank you very much for publishing my Research Article titled “Comparing Treatment Outcome Of Allergic Rhinitis Patients After Using Fluticasone Nasal Spray And Nasal Douching" in the Journal of Clinical Otorhinolaryngology. As Medical Professionals we are immensely benefited from study of various informative Articles and Papers published in this high quality Journal. I look forward to enriching my knowledge by regular study of the Journal and contribute my future work in the field of ENT through the Journal for use by the medical fraternity. The support from the Editorial office was excellent and very prompt. I also welcome the comments received from the readers of my Research Article.

img

Dr Suramya Dhamija

Dear Erica Kelsey, Editorial Coordinator of Cancer Research and Cellular Therapeutics Our team is very satisfied with the processing of our paper by your journal. That was fast, efficient, rigorous, but without unnecessary complications. We appreciated the very short time between the submission of the paper and its publication on line on your site.

img

Bruno Chauffert

I am very glad to say that the peer review process is very successful and fast and support from the Editorial Office. Therefore, I would like to continue our scientific relationship for a long time. And I especially thank you for your kindly attention towards my article. Have a good day!

img

Baheci Selen

"We recently published an article entitled “Influence of beta-Cyclodextrins upon the Degradation of Carbofuran Derivatives under Alkaline Conditions" in the Journal of “Pesticides and Biofertilizers” to show that the cyclodextrins protect the carbamates increasing their half-life time in the presence of basic conditions This will be very helpful to understand carbofuran behaviour in the analytical, agro-environmental and food areas. We greatly appreciated the interaction with the editor and the editorial team; we were particularly well accompanied during the course of the revision process, since all various steps towards publication were short and without delay".

img

Jesus Simal-Gandara

I would like to express my gratitude towards you process of article review and submission. I found this to be very fair and expedient. Your follow up has been excellent. I have many publications in national and international journal and your process has been one of the best so far. Keep up the great work.

img

Douglas Miyazaki

We are grateful for this opportunity to provide a glowing recommendation to the Journal of Psychiatry and Psychotherapy. We found that the editorial team were very supportive, helpful, kept us abreast of timelines and over all very professional in nature. The peer review process was rigorous, efficient and constructive that really enhanced our article submission. The experience with this journal remains one of our best ever and we look forward to providing future submissions in the near future.

img

Dr Griffith

I am very pleased to serve as EBM of the journal, I hope many years of my experience in stem cells can help the journal from one way or another. As we know, stem cells hold great potential for regenerative medicine, which are mostly used to promote the repair response of diseased, dysfunctional or injured tissue using stem cells or their derivatives. I think Stem Cell Research and Therapeutics International is a great platform to publish and share the understanding towards the biology and translational or clinical application of stem cells.

img

Dr Tong Ming Liu

I would like to give my testimony in the support I have got by the peer review process and to support the editorial office where they were of asset to support young author like me to be encouraged to publish their work in your respected journal and globalize and share knowledge across the globe. I really give my great gratitude to your journal and the peer review including the editorial office.

img

Husain Taha Radhi

I am delighted to publish our manuscript entitled "A Perspective on Cocaine Induced Stroke - Its Mechanisms and Management" in the Journal of Neuroscience and Neurological Surgery. The peer review process, support from the editorial office, and quality of the journal are excellent. The manuscripts published are of high quality and of excellent scientific value. I recommend this journal very much to colleagues.

img

S Munshi

Dr.Tania Muñoz, My experience as researcher and author of a review article in The Journal Clinical Cardiology and Interventions has been very enriching and stimulating. The editorial team is excellent, performs its work with absolute responsibility and delivery. They are proactive, dynamic and receptive to all proposals. Supporting at all times the vast universe of authors who choose them as an option for publication. The team of review specialists, members of the editorial board, are brilliant professionals, with remarkable performance in medical research and scientific methodology. Together they form a frontline team that consolidates the JCCI as a magnificent option for the publication and review of high-level medical articles and broad collective interest. I am honored to be able to share my review article and open to receive all your comments.

img

Tania Munoz

“The peer review process of JPMHC is quick and effective. Authors are benefited by good and professional reviewers with huge experience in the field of psychology and mental health. The support from the editorial office is very professional. People to contact to are friendly and happy to help and assist any query authors might have. Quality of the Journal is scientific and publishes ground-breaking research on mental health that is useful for other professionals in the field”.

img

George Varvatsoulias

Dear editorial department: On behalf of our team, I hereby certify the reliability and superiority of the International Journal of Clinical Case Reports and Reviews in the peer review process, editorial support, and journal quality. Firstly, the peer review process of the International Journal of Clinical Case Reports and Reviews is rigorous, fair, transparent, fast, and of high quality. The editorial department invites experts from relevant fields as anonymous reviewers to review all submitted manuscripts. These experts have rich academic backgrounds and experience, and can accurately evaluate the academic quality, originality, and suitability of manuscripts. The editorial department is committed to ensuring the rigor of the peer review process, while also making every effort to ensure a fast review cycle to meet the needs of authors and the academic community. Secondly, the editorial team of the International Journal of Clinical Case Reports and Reviews is composed of a group of senior scholars and professionals with rich experience and professional knowledge in related fields. The editorial department is committed to assisting authors in improving their manuscripts, ensuring their academic accuracy, clarity, and completeness. Editors actively collaborate with authors, providing useful suggestions and feedback to promote the improvement and development of the manuscript. We believe that the support of the editorial department is one of the key factors in ensuring the quality of the journal. Finally, the International Journal of Clinical Case Reports and Reviews is renowned for its high- quality articles and strict academic standards. The editorial department is committed to publishing innovative and academically valuable research results to promote the development and progress of related fields. The International Journal of Clinical Case Reports and Reviews is reasonably priced and ensures excellent service and quality ratio, allowing authors to obtain high-level academic publishing opportunities in an affordable manner. I hereby solemnly declare that the International Journal of Clinical Case Reports and Reviews has a high level of credibility and superiority in terms of peer review process, editorial support, reasonable fees, and journal quality. Sincerely, Rui Tao.

img

Rui Tao

Clinical Cardiology and Cardiovascular Interventions I testity the covering of the peer review process, support from the editorial office, and quality of the journal.

img

Khurram Arshad